WO2012048152A2 - Probiotic therapies for autism - Google Patents
Probiotic therapies for autism Download PDFInfo
- Publication number
- WO2012048152A2 WO2012048152A2 PCT/US2011/055159 US2011055159W WO2012048152A2 WO 2012048152 A2 WO2012048152 A2 WO 2012048152A2 US 2011055159 W US2011055159 W US 2011055159W WO 2012048152 A2 WO2012048152 A2 WO 2012048152A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- bacteria
- individual
- behavioral performance
- behavioral
- fragilis
- Prior art date
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/18—Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/20—Hypnotics; Sedatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
- A61P25/22—Anxiolytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K2035/11—Medicinal preparations comprising living procariotic cells
Definitions
- the invention relates to probiotic and probiotic-based compositions and a method of using them. More particularly, it relates to the use of Bacteroides fragilis or related bacteria to improve the mental behavior of individuals suffering from autism, autism spectrum disorders (ASD) and/or conditions with related symptoms such as anxiety.
- ASD autism spectrum disorders
- ASDs Autism spectrum disorders
- GI gastrointestinal
- a method of improving behavioral performance in an individual comprising, identifying an individual in need of treatment, and providing such an individual a composition comprising bacteria within the genus Bacteroides, whereby the individual shows improved behavioral performance.
- the individual in need of treatment suffers from anxiety, autism, ASD or a mental condition with some of the symptoms of ASD.
- the bacteria is selected from a group consisting of B. fragilis, B. theta, B. vulgatus, and B. faecalis.
- the bacteria is a mixture of several different species members within the genus Bacteroides.
- a probiotic composition for improving behavioral performance comprising bacteria within the genus Bacteroides, whereby the individual shows improved behavioral performance.
- a neutraceutical for improving behavioral performance in individuals comprising bacteria within the genus Bacteroides, whereby the individual shows improved behavioral performance.
- a pharmaceutical composition comprising bacteria within the genus Bacteroides, and a pharmaceutically acceptable carrier, whereby the composition improves behavioral performance.
- the pharmaceutical composition further comprises another pharmaceutical or compound used to treat the behavioral performance, wherein the pharmaceutical or compound does not comprise any other bacteria.
- Figure 1 shows that MIA offspring exhibit elevated anxiety in the open field test and that this can be alleviated by probiotic treatment.
- B. fragilis or other bacterial species were given in food for 1 week post-weaning (weaning is at 3 weeks of age).
- Figure 2 shows histological evidence that adult MIA offspring exhibit greater inflammatory cell infiltration of their colons.
- the inset shows a magnified view of an inflammatory cell aggregate.
- FIG 3 shows elevated IL-17 levels in lymphocytes of MIA offspring.
- MIA was induced in the pregnant mice by treatment with either poly(I:C) or a single injection of recombinant IL-6.
- the cytokine IL-6 was previously shown to induce the same ASD-like symptoms in the resulting offspring as maternal poly(I:C) injection (Smith et al., 2007).
- FIG. 4 shows that MIA offspring exhibit elevated levels of stereotyped/repetitive behavior in the marble burying tests, and that this can be normalized by probiotic treatment with certain bacteria.
- Adult offspring of saline- and poly(I:C)-injected mothers were tested for the percent of marbles buried in a 10 min period.
- B. fragilis and related species colonization at weaning prevents development of repetitive and compulsive behaviors compared to MIA offspring without the probiotic.
- Student's t-test *p ⁇ 0.05 **p ⁇ 0.01.
- n 14-20 animals per trial.
- Figure 5 shows the extent of dextran probe leakage into the circulation of MIA offspring given one of several types of bacterial species. Most important is the finding that MIA display a "leaky gut” syndrome and that B. fragilis and the other Bacteriodes species tested can prevent this disorder.
- the term "effective dose” is an amount that results in a reduction, inhibition or prevention of the behavioral disorder/abnormality/symptoms in the individual.
- the amount of B. fragilis or other probiotic required to achieve this can be determined by a person of skill in the art.
- the term "individual” as used herein includes a single biological organism wherein inflammation can occur including but not limited to animals and in particular higher animals and in particular vertebrates such as mammals and in particular human beings.
- condition/disorder/symptom or "behavioral abnormality” or as used herein means symptoms expressed by an individual with a mental disorder, such as but not limited to anxiety, autism, autism spectrum disorders, Fragile X, Rett syndrome, tuberous sclerosis, obsessive compulsive disorder, attention deficit disorder or schizophrenia.
- a mental disorder such as but not limited to anxiety, autism, autism spectrum disorders, Fragile X, Rett syndrome, tuberous sclerosis, obsessive compulsive disorder, attention deficit disorder or schizophrenia.
- the aforementioned symptoms are some of those exhibited by the MIA offspring.
- the term "individual in need of the treatment” means a person expressing or suffering from one or more of the behavioral disorder/symptoms mentioned above. An appropriately qualified person is able to identify such an individual in need of treatment using standard behavioral testing protocols/guidelines. The same behavioral testing protocols/guidelines can also be used to determine whether there is improvement to the individual's disorder/symptoms; and determine the most effective dose of the B. fragilis cell to give to an individual in need of the treatment.
- the term "improvement in behavioral performance” as used herein means a prevention or reduction in the severity or frequency, to whatever extent, of one or more of the above behavioral disorder/symptoms/abnormalities expressed by the individual. The improvement is either observed by the individual taking the treatment themselves or by another person (medical or otherwise).
- treatment indicates any activity that is part of a medical (prescribed by the physician) or non-medically approved (i.e. non-prescription including but not limited to vitamins, herbs; supplements; probiotics etc.) care that deals with a condition/symptom as described above.
- prevention indicates any activity that reduces the burden of the individual later expressing those behavioral symptoms. This takes place at primary, secondary and tertiary prevention levels, wherein: a) primary prevention avoids the development of symptoms/disorder/condition; b) secondary prevention activities are aimed at early stages of the condition/disorder/symptom treatment, thereby increasing opportunities for interventions to prevent progression of the condition/disorder/symptom and emergence of symptoms; and c) tertiary prevention reduces the negative impact of an already established
- condition/disorder/symptom by restoring function and reducing any condition/disorder/symptom or related complications.
- Pharmaceutically acceptable or appropriate carriers can be, but not limited to, organic or inorganic, solid or liquid excipient which is suitable for the selected mode of application such as oral application or injection, and administered in the form of a conventional pharmaceutical preparation.
- a conventional pharmaceutical preparation includes solid such as tablets, granules, powders, capsules, and liquid such as solution, emulsion, suspension and the like.
- Said carrier includes starch, lactose, glucose, sucrose, dextrine, cellulose, paraffin, fatty acid glyceride, water, alcohol, gum arabic and the like. If necessary, auxiliary, stabilizer, emulsifier, lubricant, binder, pH adjuster controller, isotonic agent and other conventional additives may be added.
- the pharmaceutically acceptable or appropriate carrier may well include other compounds known to be beneficial to an impaired situation of the GI tract, (e.g., antioxidants, such as Vitamin C, Vitamin E, Selenium or Zinc); or a food composition.
- the food composition can be, but is not limited to, milk, yoghurt, curd, cheese, fermented milks, milk based fermented products, ice-creams, fermented cereal based products, milk based powders, infant formulae, tablets, liquid bacterial suspensions, dried oral supplement, or wet oral supplement.
- nutraceutical means a food stuff (as a fortified food or a dietary supplement) that provides health benefits. Nutraceutical foods are not subject to the same testing and regulations as pharmaceutical drugs.
- probiotic means live microorganisms, which, when administered in adequate amounts, confer a health benefit on the host.
- the probiotics may be available in foods and dietary supplements (for example, but not limited to capsules, tablets, and powders). Examples of foods containing the probiotic are yogurt, fermented and unfermented milk, miso, tempeh, and some juices and soy beverages.
- extract indicates either the insoluble material or soluble material obtained from the B. fragilis or related species using various chemical, immunological, biochemical or physical procedures known to those of skill in the art, including but not limited to, precipitation, centrifugation, filtering, column chromatography, and detergent lysis.
- the term "whole cell lysate” refers to the fraction obtained when the B. fragilis or related species are lysed using detergent or other chemical or physical means.
- nucleic acid molecules when used in connection with biological materials such as nucleic acid molecules, polypeptides, host cells, bacterial cells/strains and the like, refers to materials as they are found in nature and not manipulated by man.
- isolated when used in connection biological materials such as nucleic acid molecules, polypeptides, bacterial cells, host cells, bacterial cells/strains and the like, refers to the isolated or purified aforementioned materials, where these materials do not occur naturally and/or where they have markedly different or distinctive characteristics compared to those found in the native material.
- non-denatured refers to when the bacterial is frozen in a media and then undergoes a freeze-drying process.
- Such non-denatured bacteria can be mixed with other substances/compounds/carriers/additives and given in forms of a pill, tablet, or liquid to individuals in need of behavioral improvement.
- the non-denatured bacteria can also be mixed into foods (cookies, yogurt; milk etc.).
- condition/disorder/symptom condition/disorder/symptom
- condition/behavioral abnormality
- the Patterson laboratory developed a mouse model that has both construct and face validity for autism, MIA (maternal immune activation ) (reviewed in Patterson, 2011). This is based on epidemiological evidence that viral or bacterial infection during pregnancy increases the risk for ASD in the offspring (Atladottir et al., 2010). In this animal model, pregnant mice that receive a respiratory infection at mid-gestation, or that receive the double-stranded RNA viral mimic, poly(I:C), produce offspring with a series of abnormal behaviors, including the hallmark symptoms of repetitive/compulsive behaviors, and deficits in social interaction and communication (Patterson, 2011).
- MIA offspring display a specific neuropathology that is common in ASD, spatially restricted deficits in Purkinje cells (Patterson, 2011). Recently, the collaboration between the laboratories of Sarkis Mazmanian and Paul Patterson has discovered that the MIA offspring exhibit abnormalities in the immune system and the gastrointestinal tract. Most importantly, some of the ASD-like behavioral symptoms can be corrected or prevented by manipulating the microbiota of the MIA offspring. This treatment is based on prior work by the Mazmanian group showing the efficacy of B. fragilis treatment in several GI and systemic inflammatory disorders (Round et al., 2009, 2010).
- the open field test is a test of anxiety under mildly stressful conditions, and ASD subjects exhibit enhanced anxiety under such circumstances.
- the offspring are tested in the open field to determine the effects of B. fragilis and a number of other bacterial species on anxiety behavior.
- Groups of pregnant mice (n>3) are treated on El 2.5 with poly(I:C) to induce MIA, or saline as the control (Smith et al., 2007).
- Immediately upon weaning the offspring are colonized with one of the bacteria shown (given in food) or not, for one week. Mice are placed in a brightly lit open box, and activity is recorded by a video camera and analyzed using Ethovision software (Noldus).
- MIA offspring spend less time in the center of the open field and enter it less often than control offspring, indicating increased anxiety (Fig. 1).
- Treatment with B. fragilis and other Bacteroides species prevents the abnormal behavior in the MIA offspring, indicating that this probiotic treatment lowers anxiety in normal animals as well as in the ASD model mice.
- the non-Bacteroides species, E.rioselis is not able to prevent anxiety in the MIA offspring.
- lymphocytes associated with the GI tract mesenteric lymph nodes were dissected from adult offspring of control mothers, offspring of MIA mothers treated with poly(I:C), and offspring of MIA mothers treated with IL-6.
- CD4+ cells were isolated and treated in culture with PMA and ionomycin. Supematants were assayed for IL-6 (not shown) and IL-17 (Fig. 3). On both days assayed, the lymphocytes from MIA offspring display higher cytokine release than controls.
- GI epithelial barrier function can be tested by gavaging the mouse with a labeled dextran that is large enough such that it does not normally leak through the barrier in significant amounts.
- DSS dextran sodium sulfate
- the labeled dextran probe does leak into the circulation (Fig. 5). This is the positive control for "leaky gut", a symptom found in a large subset of ASD children.
- the adult MIA offspring also display very significant levels of leaky gut, and this can be prevented by post- weaning treatment with all of the Bacteroides species tested, but not with the non-Bacteroides, E. faecalis (Fig. 5).
- the efficacy of probiotic therapy in the GI tract corresponds with the specificity of the behavioral therapy.
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- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Anesthesiology (AREA)
- Psychiatry (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
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Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ES11831629.8T ES2581843T3 (es) | 2010-10-07 | 2011-10-06 | Terapias probióticas para el autismo |
JP2013532955A JP5931884B2 (ja) | 2010-10-07 | 2011-10-06 | 自閉症の生菌療法 |
CA2813606A CA2813606C (en) | 2010-10-07 | 2011-10-06 | Probiotic therapies for autism |
EP11831629.8A EP2624863B1 (en) | 2010-10-07 | 2011-10-06 | Probiotic therapies for autism |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US39100410P | 2010-10-07 | 2010-10-07 | |
US61/391,004 | 2010-10-07 | ||
US201161472963P | 2011-04-07 | 2011-04-07 | |
US61/472,963 | 2011-04-07 |
Publications (2)
Publication Number | Publication Date |
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WO2012048152A2 true WO2012048152A2 (en) | 2012-04-12 |
WO2012048152A3 WO2012048152A3 (en) | 2012-06-21 |
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PCT/US2011/055159 WO2012048152A2 (en) | 2010-10-07 | 2011-10-06 | Probiotic therapies for autism |
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US (3) | US9452189B2 (sl) |
EP (2) | EP3072524B1 (sl) |
JP (2) | JP5931884B2 (sl) |
CA (1) | CA2813606C (sl) |
CY (1) | CY1120119T1 (sl) |
DK (1) | DK3072524T3 (sl) |
ES (2) | ES2581843T3 (sl) |
HR (1) | HRP20180559T1 (sl) |
HU (1) | HUE038600T2 (sl) |
LT (1) | LT3072524T (sl) |
PL (1) | PL3072524T3 (sl) |
PT (1) | PT3072524T (sl) |
RS (1) | RS57006B1 (sl) |
SI (1) | SI3072524T1 (sl) |
WO (1) | WO2012048152A2 (sl) |
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Also Published As
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JP5931884B2 (ja) | 2016-06-08 |
CY1120119T1 (el) | 2018-12-12 |
PL3072524T3 (pl) | 2018-06-29 |
HRP20180559T1 (hr) | 2018-06-01 |
PT3072524T (pt) | 2018-04-23 |
US20120087895A1 (en) | 2012-04-12 |
US20210000888A1 (en) | 2021-01-07 |
WO2012048152A3 (en) | 2012-06-21 |
ES2581843T3 (es) | 2016-09-07 |
JP2013544780A (ja) | 2013-12-19 |
DK3072524T3 (en) | 2018-04-09 |
HUE038600T2 (hu) | 2018-10-29 |
JP2016185959A (ja) | 2016-10-27 |
ES2662412T3 (es) | 2018-04-06 |
EP2624863A4 (en) | 2014-04-02 |
LT3072524T (lt) | 2018-03-26 |
CA2813606A1 (en) | 2012-04-12 |
EP3072524B1 (en) | 2018-01-17 |
JP6177379B2 (ja) | 2017-08-09 |
RS57006B1 (sr) | 2018-05-31 |
EP2624863A2 (en) | 2013-08-14 |
US20160375065A1 (en) | 2016-12-29 |
US9452189B2 (en) | 2016-09-27 |
EP2624863B1 (en) | 2016-04-20 |
SI3072524T1 (sl) | 2018-04-30 |
CA2813606C (en) | 2019-01-22 |
US11896629B2 (en) | 2024-02-13 |
EP3072524A1 (en) | 2016-09-28 |
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